Encapsulation, stabilization, and release of BSA-FITC from polyanhydride microspheres
Encapsulation, stabilization, and release of BSA-FITC from polyanhydride microspheres
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DOI：
10.1016/j.jconrel.2004.08.006
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发表时间：
2004-11-05
影响因子：
10.8
通讯作者：
Narasimhan, B
中科院分区：
文献类型：
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作者：
Determan, AS;Trewyn, BG;Narasimhan, B
为了确定多酸酐作为治疗性蛋白载体的效果，我们将用异硫氰酸荧光素（BSA-FITC）标记的模型蛋白牛血清白蛋白包被在聚己二酸酐（聚（SA））和聚（1,6-双对羧基苯氧基）己烷（聚（CPH））的随机共聚物微球中。通过双乳液（水/油/水）技术制备微球，并使用扫描电镜、凝胶渗透色谱、共聚焦显微镜和Coulter计数器对微球进行了表征。在体外释放研究中考察了蛋白质负载、蛋白质分布和聚合物组成变化的影响。用傅里叶变换红外光谱测定了包封BSA-FITC的二级结构。用十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析了释放蛋白的一级结构。Poly（SA）和20:80 （CPH:SA）微球保存了释放蛋白的一级结构和被封装蛋白的二级结构，并分别表现出约15和30天的持续递送。随着共聚物中CPH含量的增加，FITC-BSA的二级结构不再保守，α -螺旋含量急剧下降。(C) 2004 Elsevier B.V.版权所有
In order to determine the efficacy of using polyanhydrides as a carrier for therapeutic proteins, the model protein bovine serum albumin labeled with fluorescein isothiocyanate (BSA-FITC) was encapsulated in microspheres of poly sebacic anhydride (poly(SA)), and random copolymers of poly(SA) and poly(1,6-bis-p-carboxyphenoxy)hexane (poly(CPH)). The microspheres were fabricated via the double emulsion (water/oil/water) technique and were characterized using scanning electron microscopy, gel permeation chromatography, confocal microscopy, and a Coulter counter. The effect of protein loading, protein distribution, and change in polymer composition was examined in an in vitro release study. The secondary structure of the encapsulated BSA-FITC was determined with Fourier transform infrared spectroscopy. The primary structure of the released protein was analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Poly(SA) and 20:80 (CPH:SA) microspheres were found to conserve the primary structure of the released protein and the secondary structure of the encapsulated protein, and showed a sustained delivery for approximately 15 and 30 days, respectively. As the CPH content in the copolymer increased, the secondary structure of FITC-BSA was not conserved, as indicated by the steep decrease in the alpha-helix content. (C) 2004 Elsevier B.V. All rights reserved.